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Molecular prevalence and characterization of Cryptosporidium in domestic free-range poultry in Anhui Province, China

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Zheng Gong
Zheng Gong
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Zhen-zhen Kan
Zhen-zhen Kan
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Jia-min Huang
Jia-min Huang
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Free-range chickens might mediate the spread of Cryptosporidium oocysts to humans and other animals. Few studies have evaluated the prevalence of Cryptosporidium species in domestic free-range poultry in China. Here, we characterized the prevalence and distribution of species and genotypes of Cryptosporidium in domestic free-range chickens, ducks, and geese in Anhui Province, China. A total of 1910 fresh fecal samples from three poultry species were examined from 18 free-range poultry farms by nested PCR and analysis of the Cryptosporidium SSU rRNA gene. The overall prevalence of Cryptosporidium species was 2.9% (55/1910), with infection rates of 1.3% (11/829) in chickens, 7.3% (36/487) in ducks, and 1.4% (8/594) in geese. C. baileyi (0.6%), C. meleagridis (0.2%), C. galli (0.2%), and C. xiaoi-like genotype (0.2%) were identified in chickens, and only C. baileyi was identified in ducks and geese, with infection rates of 7.4% and 1.3%, respectively. C. baileyi was the most prevalent species. Sequencing of the GP60 gene revealed that the C. meleagridis isolates belonged to the IIIbA26G1R1b subtype. This is the first study to document C. galli and C. xiaoi-like genotype in domestic free-range chickens in China. These findings expand the range of avian hosts known for Cryptosporidium and highlight the need for additional studies to characterize the diversity of Cryptosporidium in avian species.
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https://doi.org/10.1007/s00436-021-07191-y
PROTOZOOLOGY - ORIGINAL PAPER
Molecular prevalence andcharacterization ofCryptosporidium
indomestic free‑range poultry inAnhui Province, China
ZhengGong1· Zhen‑zhenKan1· Jia‑minHuang1· ZhuiFang1· Xin‑chaoLiu1· You‑fangGu1· Wen‑ChaoLi1
Received: 1 April 2021 / Accepted: 11 May 2021
© The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature 2021
Abstract
Free-range chickens might mediate the spread of Cryptosporidium oocysts to humans and other animals. Few studies have
evaluated the prevalence of Cryptosporidium species in domestic free-range poultry in China. Here, we characterized the
prevalence and distribution of species and genotypes of Cryptosporidium in domestic free-range chickens, ducks, and geese
in Anhui Province, China. A total of 1910 fresh fecal samples from three poultry species were examined from 18 free-range
poultry farms by nested PCR and analysis of the Cryptosporidium SSU rRNA gene. The overall prevalence of Crypto-
sporidium species was 2.9% (55/1910), with infection rates of 1.3% (11/829) in chickens, 7.3% (36/487) in ducks, and 1.4%
(8/594) in geese. C. baileyi (0.6%), C. meleagridis (0.2%), C. galli (0.2%), and C. xiaoi-like genotype (0.2%) were identified
in chickens, and only C. baileyi was identified in ducks and geese, with infection rates of 7.4% and 1.3%, respectively. C.
baileyi was the most prevalent species. Sequencing of the GP60 gene revealed that the C. meleagridis isolates belonged to
the IIIbA26G1R1b subtype. This is the first study to document C. galli and C. xiaoi-like genotype in domestic free-range
chickens in China. These findings expand the range of avian hosts known for Cryptosporidium and highlight the need for
additional studies to characterize the diversity of Cryptosporidium in avian species.
Keywords Prevalence· C. baileyi· C. meleagridis· C. galli· C. xiaoi-like genotype
Introduction
Intracellular parasites of the genus Cryptosporidium (Api-
complexa: Cryptosporidiidae) are a common cause of gas-
trointestinal diseases in vertebrates and respiratory diseases
in birds (Nakamura and Meireles 2015; Wang etal. 2019).
Over 40 species and more than 70 genotypes of Crypto-
sporidium have been identified to date based on molecular
and morphological data (Feng 2018; Wang etal. 2019; Innes
etal. 2020). Infections in birds are mainly caused by four
species: C. baileyi, C. meleagridis, C. galli, and C. avium
(previously known as avian genotype V) (Nakamura and
Meireles, 2015; Wang etal. 2019). C. baileyi is generally
the most prevalent in domestic poultry, and C. meleagridis is
the only avian Cryptosporidium species with zoonotic poten-
tial (Helmy etal. 2017; Santana etal. 2018). In addition, 21
avian Cryptosporidium genotypes, including Cryptosporid-
ium goose genotypes (I–IV), a Cryptosporidium duck gen-
otype, and Cryptosporidium avian genotypes (I–IV), have
been identified in more than 30 avian species worldwide,
and several mammal-specific Cryptosporidium species and
genotypes, including C. andersoni, C. hominis, C. muris, C.
parvum, C. canis, and muskrat genotype I, have been infre-
quently reported in birds (Ryan 2010; Holubová etal. 2016,
2019; Wang etal. 2019).
Although cryptosporidiosis is one of the most common
parasitic infections in poultry worldwide, literature related
to the occurrence of Cryptosporidium infection in avian spe-
cies is scarce, especially compared with mammals (Elkarim
Laatamna etal. 2017). Although some studies have reported
Cryptosporidium infections in domestic chickens, ducks,
and geese in China, most Cryptosporidium infections in
early studies were identified on the basis of the presence
Section Editor: Yaoyu Feng
Zheng Gong, Zhen-zhen Kan, Jia-min Huang, and Zhui Fang
contributed equally to this work.
* Wen-Chao Li
liwen303@126.com
1 Anhui Province Key Laboratory ofAnimal Nutritional
Regulation andHealth, College ofAnimal Science, Anhui
Science andTechnology University, Fengyang233100,
People’sRepublicofChina
/ Published online: 21 August 2021
Parasitology Research (2021) 120:3519–3527
Content courtesy of Springer Nature, terms of use apply. Rights reserved.
... Cryptosporidiosis is a zoonotic protozoan infection among wide range of hosts including birds which is considered as a problematic state to poultry industry worldwide. Due to the lack of the study regarding the importance of cryptosporidiosis in poultry as reservoir, the current study detects Cryptosporidium parasite through microscopy as preliminarily effort, with the overall infection rate of 29.30%, such high prevalence was disagreed with the reported lower infection rate of 2.9% by Gong et al. [27]. ...
... The finding of AL-Zubaidi et al. [28] in broiler 35% somewhat harmony with current data 38%. In different to the study data lower prevalence of 1.3% was reported by [27], although 12.6% and 13.2% were reported by some investigators [29] and [30] from free range and broiler chicken respectively. Contrary to the current prevalence of cryptosporidiosis in pigeon 35.56% lower prevalence of 20% and 21% were reported through applying of modified acid fast, and PCR by some authors [31,32]. ...
... Regarding other inspected poultry species, current data represent the occurrence of cryptosporidiosis in geese and ducks with a prevalence rate of 24.44% and 22.5% respectively. Contrary to the current data [27] reported a low prevalence of 7.3% and 1.4% in both ducks and geese respectively. Besides that, higher prevalence of 39.9% and 50% was reported in ducks by some researchers [34,35]. ...
Detection of Natural Infection with Cryptosporidiosis in Domestic Poultry from Sulaymaniyah Province/ Iraq
Article
Full-text available
  • Jan 2024
CRYPTOSPORIDIOSIS is Cryptosporidiosis is a zoonotic protozoan infection caused by Cryptosporidium spp., with a public health concern. It has been reported widely across several countries in both wild and domestic birds. The study was designed to find out the Cryptosporidium parasite in different species of domestic poultry from Sulaymaniyah province northern region of Iraq. The study was carried during the period between June 2022 to February 2023, a total of 215 fresh dropping samples were collected from chicken, turkey, duck, geese and pigeons. All samples were screened for the presence of Cryptosporidium parasite by microscopy using fecal flotation and modified Zeihl- Neelsen staining method for recognition the oocyst stage of the parasite. Intestinal tissue samples were also fixed in 10% buffered formalin and stained with (H&E) for histopathology, to find out the lesions. The overall infection rate of Cryptosporidiosis was 29.30% among the examined poultry species, with the higher prevalence of 38.00% in chicken, followed by pigeon 35.56% then in geese 24.44%, and turkey 22.86%, and a lower prevalence of 22.50% was reported in duck. Histopathology from intestinal tissue revealed sever epithelial damage, ulceration with sloughing of villi, accompanied by infiltration of inflammatory cells with extensive edema. The results of the present study provide the first data on the occurrence of Cryptosporidium infection from domestic poultry in the study area.
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... A total of 1910 fresh fecal samples from free-range chickens (n = 829), ducks (n = 487), and geese (n = 594) were collected from 18 different domestic free-range poultry farms in Anhui Province as described previously by Gong et al (2021) (Fig. 1 and Table 1). ...
... Regardless of the relative lack of importance of T. gallinarum in the poultry industry, the low infection rates of T. gallinarum in this study are in accordance with the current, but sparse, published information regarding the global epidemiology of T. gallinarum from chickens, ducks, and geese. The low occurrence of T. gallinarum in this study may be attributed mainly to the fecal specimen process (Gong et al. 2021). In the present study, genomic DNA was extracted directly from untreated fecal samples, but the DNA was extracted from the culture of positive clinical specimens with more live organisms in other studies (Badparva et al. 2020;Carrisosa et al. 2021). ...
... Considering the presence of inhibitors in extracted feces, genomic DNAs tend to reduce the sensitivity of PCR detection; the Stool DNA Kits used in different laboratories may be also a factor involved in the difference of infection rates. However, prevalence data can be influenced by many factors, including the different detection methods used in studies (such as microscopic examination and PCR), sample origin (tissue samples vs. feces), sample size, animal management, animal health, hygiene, and biosecurity conditions (Gong et al. 2021). Further studies are needed to clarify this issue. ...
Molecular prevalence of Tetratrichomonas gallinarum and Trichomonas gallinae in three domestic free-range poultry breeds in Anhui Province, China
Article
Full-text available
  • Aug 2022
  • Parasitol Res
Tetratrichomonasgallinarum and Trichomonas gallinae can colonize the alimentary tract of domestic birds. However, little information is available on the epidemiology of the two trichomonad species in domestic free-range poultry in China. In this study, the occurrence and genetic characteristic of T. gallinarum and T. gallinae among free-range chickens, ducks, and geese in Anhui Province, China, were investigated. The 1910 fecal samples collected from 18 free-range poultry farms throughout Anhui Province were examined for the presence of T. gallinarum and T. gallinae by PCR and sequence analysis of the small subunit (SSU) rRNA gene of T. gallinarum and ITS1-5.8S-ITS2 sequence of T. gallinae. The overall occurrence of T. gallinarum in poultry was 1.2% (22/1910), with infection rates of 2.1% (17/829) in chickens, 0.2% (1/487) in ducks, and 0.7% (4/594) in geese. The constructed phylogeny tree using the concatenated ITS1-5.8S-ITS2 region and SSU rRNA indicated the T. gallinarum isolates detected in this study were closely related to previously defined genogroups A, D, and E, respectively. Nine (0.5%) fecal samples were positive for T. gallinae, with infection rates of 0.8% (7/829) in chickens, 0.4% (2/487) in ducks, and 0% (0/594) in geese. Sequence and phylogenetic analysis showed that four T. gallinae ITS1-5.8S-ITS2 sequences obtained from chicken feces and one duck fecal sample belonged to genotype ITS-OBT-Tg-1. This is the first report of the prevalence and genetic characterization of T. gallinarum and T. gallinae in free-range chickens, ducks, and geese in China.
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... Cryptosporidium meleagridis has been described in a wide variety of avian species (Table 1). The majority of reports in birds have been in farmed and caged birds or birds in rehabilitation centres, probably due to the high density and overcrowding of susceptible animals (Nakamura et al., 2009, Abe andWang et al., 2010;Berrilli et al., 2012;Silverlås et al., 2012;Wang et al., 2012;Baroudi et al., 2013;Wang et al., 2014;Li et al., 2015;Reboredo-Fernández et al., 2015, Máca and Pavlásek, 2016, Ewald et al., 2017Ayinmode et al., 2018;Da Cunha et al., 2018;Liao et al., 2018;Santana et al., 2018;Kabir et al., 2020;Dong et al., 2021;Gong et al., 2021;Liao et al., 2021;Feng et al., 2022aFeng et al., , 2022bLin et al., 2022). ...
The risk of wild birds contaminating source water with zoonotic Cryptosporidium and Giardia is probably overestimated
Article
Full-text available
  • Dec 2023
  • SCI TOTAL ENVIRON
Cryptosporidium and Giardia are important waterborne protozoan parasites that are resistant to disinfectants commonly used for drinking water. Wild birds, especially wild migratory birds, are often implicated in the contamination of source and wastewater with zoonotic diseases, due to their abundance near water and in urban areas and their ability to spread enteric pathogens over long distances. This review summarises the diversity of Cryptosporidium and Giardia in birds, with a focus on zoonotic species, particularly in wild and migratory birds, which is critical for understanding zoonotic risks. The analysis revealed that both avian-adapted and zoonotic Cryptosporidium species have been identified in birds but that avian-adapted Cryptosporidium species dominate in wild migratory birds. Few studies have examined Giardia species and assemblages in birds, but the non-zoonotic Giardia psittaci and Giardia ardeae are the most commonly reported species. The identification of zoonotic Cryptosporidium and Giardia in birds, particularly C. parvum and G. duodenalis assemblages A and B in wild migratory birds, is likely due to mechanical carriage or spillback from birds co-grazing pastures contaminated with C. parvum from livestock. Therefore, the role of wild migratory birds in the transmission of zoonotic Cryptosporidium and Giardia to source water is likely overestimated. To address knowledge gaps, it is important to conduct more extensive studies on the prevalence of Cryptosporidium and Giardia in a broader range of migratory wild birds. There is also a need to investigate the extent to which zoonotic infections with C. hominis/C. parvum and G. duodenalis assemblages A and B are mechanical and/or transient, and to assess the load and viability of zoonotic oo/cysts shed in avian faeces. Understanding the contribution of birds to zoonoses is essential for effective disease surveillance, prevention, and control.
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... In Merauke Regency, the population of free-range chickens is approximately 1,682,728 (BPS, Merauke). The growth of free-range differs from purebred and broiler chickens (Aksoy, et al, 2021), particularly due to their relatively slower growth (Casartelli, et al, 20216), ( Gong et al, 2021). The critical factors influencing production are feed and genetics. ...
The Contribution of Agricultural Waste to the Production Performance of Free-Range Chickens in the Easternmost Region of Indonesia
Article
Full-text available
  • Oct 2023
Purpose: This research aims to investigate the production performance of free-range chickens fed with banana peel flour as a substitute for bran in the ration. Theoretical framework: Free-range chickens, commonly known as "non-purebred chickens", are closely related to the community. These chickens exhibit diverse genetic traits and are commonly found in the wider community, for both animal protein consumption and as a supplementary source of income to improve their economy. Design/methodology/approach: The research was conducted in Merauke from October to December 2022, and the materials used included 48 three-week-old free-range chickens with a weight of 150-200g, concentrate, yellow corn, bran, and banana peel flour. A Randomized Block Design was used with 4 treatments namely P0 = control diet (0% banana peel flour), as well as P1 = 25%, P2 = 50%, and P3 = 100% banana peel flour substituting bran, distributed into 4 groups based on body weight. The observed variables were ration consumption, weight gain, and ration conversion. Meanwhile, data were analyzed using analysis of variance and further tested with the least significant difference (LSD) test Findings: The results showed that the use of banana peel flour as a substitute for bran significantly reduced ration consumption (P<0.05) but did not have a significant effect (P>0.05) on weight gain and ration conversion. Research, Practical & Social implications: Utilization of banana peel waste as animal feed will have an impact on reducing environmental pollution and changing the socio-economic conditions of farmers. Originality/value:Research using banana peel waste as poultry feed is the first time it has been conducted in Merauke Regency
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... Based on molecular characterization of the SSU rRNA gene, two species, C. meleagridis and C. baileyi, were determined in this study, which were also observed as the common Cryptosporidium species in chickens in other provinces of China (Table 3) and in Algeria and Sweden (Baroudi et al. 2013;Cao et al. 2020;Gong et al. 2021;Laatamna et al. 2018;Liao et al. 2018;Silverlas et al. 2012). Surprisingly, one of C. meleagridis and C. baileyi was found, or other species and genotypes were confirmed simultaneously in chickens in most countries. ...
Dominant infection of Cryptosporidium baileyi in broiler chickens in Zhejiang Province, China
Article
Full-text available
  • Jun 2023
  • Parasitol Res
Cryptosporidium is a common enteric parasite in chickens. A total of 812 fecal specimens were collected from 11 broiler farms in Zhejiang Province, China, and analyzed by nested PCR amplification based on the small subunit ribosomal RNA (SSU rRNA) gene. The overall infection rate of Cryptosporidium was 6.3% (51/812), and five of 11 farms were Cryptosporidium positive. Broilers aged > 90 days accounted for the highest infection rate of 16.1% (6/56), followed by those aged 30–60 days (10.6%, 38/358) and 60–90 days (4/378, 1.1%). Two Cryptosporidium species were identified by sequence analysis, with the predominant species being C. baileyi (96.1%, 49/51) and the minor infection being C. meleagridis (3.9%, 2/51). Based on the 60-kDa glycoprotein (gp60) gene, two C. meleagridis-positive isolates were identified as one known subtype, IIIbA24G1R1. This study indicated the common occurrence of C. baileyi in broiler chickens in this region and low zoonotic transmission potential of Cryptosporidium to humans.
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... To our knowledge, this is the first report on the presence and prevalence of Cryptosporidium in intensively farmed chickens in Guangdong Province, although previous studies have been reported in Hubei, Zhejiang, Henan and Anhui in China [3,5,8,25]. In our study, the overall prevalence of Cryptosporidium in chickens (13.2%; 132/1001) was comparable to previous numbers reported for domestic chickens in Brazil (12.6%) [10], China (10.2%) [8] and Syria (9.9%) [26], and higher than Iran (0.5%) [27], Tunisia (4.5%) [28], Jordan (4.8%) [29] and Germany (5.7%) [15], but lower than Brazil (25.6%) [30] and Algeria (34.4%) [4]. ...
Dominance of the zoonotic pathogen Cryptosporidium meleagridis in broiler chickens in Guangdong, China, reveals evidence of cross-transmission
Article
Full-text available
  • Jun 2022
Background Cryptosporidium is one of the most prevalent parasites infecting both birds and mammals. To examine the prevalence of Cryptosporidium species and evaluate the public health significance of domestic chickens in Guangdong Province, southern China, we analyzed 1001 fecal samples from 43 intensive broiler chicken farms across six distinct geographical regions. Methods Individual DNA samples were subjected to nested PCR-based amplification and sequencing of the small subunit of the nuclear ribosomal RNA gene ( SSU rRNA). Analysis of the 60 kDa glycoprotein gene ( gp 60) was performed to characterize the subtypes of C. meleagridis . Results The overall prevalence of Cryptosporidium was 13.2% (95% CI 11.1–15.3) (24 of 43 farms), with C. meleagridis (7.8%), C. baileyi (4.8%) and mixed infections (0.6%). Using the gp 60 gene, three subtype families, IIIb, IIIe and IIIg, were identified, including six subtypes: one novel (IIIgA25G3R1a) and five previously reported (IIIbA23G1R1c, IIIbA24G1R1, IIIbA21G1R1a, IIIeA17G2R1 and IIIeA26G2R1). Within these subtypes, five known subtypes were genetically identical to those identified in humans. Conclusions This is the first report of C. meleagridis in chickens from Guangdong. The frequent occurrence of C. meleagridis in domestic chickens and the common C. meleagridis subtypes identified in both humans and chickens is of public health significance. Our study indicates that broiler chickens represent a potential zoonotic risk for the transmission of Cryptosporidium in this region. Graphical Abstract
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... Six Cryptosporidium species infect birds, namely, C. meleagridis (Slavin, 1955), C. baileyi , C. galli (Ryan et al., 2003), C. avium (avian genotype V) (Holubová et al., 2016), C. proventriculi (avian genotype III) Holubová et al., 2019), and C. ornithophilus (avian genotype II) (Santos et al., 2005;Meireles et al., 2006;Ng et al., 2006;Holubová et al., 2020). Cryptosporidium avian genotypes I, IV, VI, VII, VIII, and IX (Ng et al., 2006;Helmy et al., 2017) and Cryptosporidium xiaoi-like genotype (Ewald et al., 2017;Santana et al., 2018;Liu et al. 2019;Gong et al., 2021) have also been described in birds. ...
Validation of a one-tube nested real-time PCR assay for the detection of Cryptosporidium spp. in avian fecal samples
Article
Full-text available
  • Mar 2022
  • REV BRAS PARASITOL V
The aim of this study was to validate a one-tube nested real-time PCR assay followed by genetic sequencing to detect and identify Cryptosporidium species and genotypes in birds. A total of 443 genomic DNA extracted from avian fecal samples were analyzed by one-tube nested real-time PCR and conventional nested PCR. By one-tube nested real-time PCR, 90/443 (20.3%) samples were positive for Cryptosporidium spp. In contrast, 36/443 (8.1%) samples were positive for Cryptosporidium spp. by conventional nested PCR. The analytical sensitivity test showed that one-tube nested real-time PCR detects approximately 0.5 oocyst (2 sporozoites) per reaction. An evaluation of analytical specificity did not reveal amplification of microorganisms that commonly present nonspecific amplification with primers used for the diagnosis of Cryptosporidium spp. The repeatability analysis showed the same result in 27 out of 30 samples (90%). As for the reproducibility of one-tube nested real-time PCR, 24 of the 30 samples examined (80%) showed the same result. All the 90 samples amplified by one-tube real-time nested PCR were successfully sequenced, leading to the identification of C. baileyi, C. galli, C. meleagridis, C. proventriculi, and Cryptosporidium avian genotype I. Genetic sequencing of conventional nested PCR amplicons was successful in 10/36 (27.8%) of positive samples.
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... To our knowledge, this is the rst report of the presence and prevalence of Cryptosporidium in intensively farmed chickens in Guangdong Province, although previous studies have reported in Hubei, Zhejiang, Henan and Anhui in China [4,9,18,19]. In our study, the overvall prevalence of Cryptosporidium in chickens (13.2%; 132/1001) was comparable to previous values reported for domestic chickens in Brazil (12.6%; 24/190) [20], China (10.2%) [9], Syria (9.9%) [21], higher than Iran (0.5%) [22], Tunisia (4.5%) [23], Jordan (4.8%) [24], and Germany (5.7%) [13], but lower than Brazil (25.6%) [25] and Algeria (34.4%) [5]. ...
Dominance of Zoonotic Pathogen Cryptosporidium Meleagridis in Broiler Chickens in Guangdong, China, Reveals Evidence of Cross-Transmission
Preprint
Full-text available
  • Nov 2021
Background Cryptosporidium is one of the most prevalent parasites infecting both birds and mammals. To examine the prevalence of Cryptosporidium species and evaluate the public health significance of domestic chickens in Guangdong Province, Southern China, we analyzed 1001 fecal samples collected from 43 intensive broiler chicken farms from six distinct geographical regions between June 2020 and March 2021. Methods Individual DNAs were subjected to nested PCR-based amplification and sequencing of the small subunit of the nuclear ribosomal RNA gene (SSU rRNA). The 60 kDa glycoprotein gene (pgp60) was performed from all positive SSU rRNA samples to characterise subtypes of C. meleagridis. Results Cryptosporidium infection rates was found to be 13.2%, comprising with infections with C. meleagridis (78/1001, 7.8%), C. baileyi (48/1001, 4.8%) and mixed infections (6/1001, 0.6%). Three subtype families were identified, IIIb, IIIe and IIIg. Six subtypes were identified in broiler chickens, including one novel (IIIgA25G3R1a) and five previously reported (IIIbA23G1R1c, IIIbA24G1R1, IIIbA21G1R1a, IIIeA17G2R1 and IIIeA26G2R1). Within these subtypes, five known subtypes were genetically identical to those identified in humans. Conclusions This is the first report of C. meleagridis in chickens from Guangdong. The frequent occurrence of C. meleagridis in domestic chickens and the common C. meleagridis subtypes identified both in humans and chickens is of public health significance. Our study indicates that broiler chickens represent a potential zoonotic risk for the transmission of Cryptosporidium in this region.
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Genetic characterization of Cryptosporidium spp. in Hotan Black Chickens in China reveals two novel subtypes of Cryptosporidium meleagridis
Article
Full-text available
  • Nov 2022
  • Parasite
A total of 617 fecal specimens were collected on 18 Hotan Black chicken farms in Southern Xinjiang, China, and tested for the presence of Cryptosporidium spp. by PCR of the small subunit ribosomal RNA (SSU rRNA) gene. The overall infection rate by Cryptosporidium spp. was 11.5% (71/617), and ten of the 18 farms were positive. The infection rate by Cryptosporidium spp. was 14.5% (48/331) in the 30–60 d group, higher than chickens in the <30 d (12.0%, 15/125), 60–90 d (6.9%, 5/72), and >90 d (3.4%, 3/89) groups. Cryptosporidium meleagridis (n = 38) and C. baileyi (n = 33) were confirmed by sequencing analysis. A total of 25 of the 38 C. meleagridis-positive specimens were subtyped successfully at the gp60 gene, including one known subtype (IIIbA23G1R1, n = 1) and two novel subtypes, named IIIbA25G1R1 (n = 20) and IIIbA31G1R1 (n = 4). The results showed that infection by Cryptosporidium spp. in Hotan Black Chickens was common in this area and the distribution of C. meleagridis subtypes had regional characteristics.
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Longitudinal surveillance of Cryptosporidium spp. in broiler chickens in Xinjiang, northwest China: genetic diversity of Cryptosporidium meleagridis subtypes
Article
Full-text available
  • Oct 2022
  • Parasitol Res
Cryptosporidium spp. are common enteric parasites in humans and animals. Herein, 175 faecal specimens were collected from a broiler farm in Xinjiang, China, including seven repeated samplings at 10-day intervals of broilers aged 10 to 70 days. Cryptosporidium was detected and identified by PCR–RFLP analysis. The overall infection rate of Cryptosporidium in broilers was 23.4% (41/175), with the highest infection rate of 48.0% (12/25) at 40 days of age, and no infection was detected at 10 days of age. Two Cryptosporidium species were confirmed, namely, C. baileyi (3.4%, 6/175) and C. meleagridis (20%, 35/175). In total, 21 of 35 C. meleagridis isolates were successfully subtyped based on the gp60 gene, and one known subtype, IIIgA22G3R1 (n = 1), and three novel subtypes, IIIbA25G1R1 (n = 10), IIIgA24G3R1 (n = 9) and IIIgA25G2R1 (n = 1), were identified. Our findings highlight the genetic diversity of C. meleagridis in Xinjiang and the potential endemic characteristics of the subtypes.
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A retrospective epidemiological analysis of human Cryptosporidium infection in China during the past three decades (1987-2018)
Article
Full-text available
Background Cryptosporidiosis is an emerging infectious disease of public health significance worldwide. The burden of disease caused by Cryptosporidium varies between and within countries/areas. To have a comprehensive understanding of epidemiological status and characteristics of human Cryptosporidium infection in China since the first report in 1987, a retrospective epidemiological analysis was conducted by presenting differences in the prevalence of Cryptosporidium by province, year, population, living environment and season and possible transmission routes and risk factors as well as genetic characteristics of Cryptosporidium in humans. Methodology/Principal findings A systematic search was conducted to obtain epidemiological papers of human Cryptosporidium infection/cryptosporidiosis from PubMed and Chinese databases. Finally, 164 papers were included in our analysis. At least 200,054 people from 27 provinces were involved in investigational studies of Cryptosporidium, with an average prevalence of 2.97%. The prevalence changed slightly over time. Variable prevalences were observed: 0.65–11.15% by province, 1.89–47.79% by population, 1.77–12.87% and 0–3.70% in rural and urban areas, respectively. The prevalence peak occurred in summer or autumn. Indirect person-to-person transmission was documented in one outbreak of cryptosporidiosis in a pediatric hospital. 263 Cryptosporidium isolates were obtained, and seven Cryptosporidium species were identified: C. hominis (48.3%), C. andersoni (22.43%), C. parvum (16.7%), C. meleagridis (8.36%), C. felis (3.04%), C. canis (0.76%) and C. suis (0.38%). Conclusions/Significances This systematic review reflects current epidemiological status and characteristics of Cryptosporidium in humans in China. These data will be helpful to develop efficient control strategies to intervene with and prevent occurrence of human Cryptosporidium infection/cryptosporidiosis in China as well as have a reference effect to other countries. Further studies should focus on addressing a high frequency of C. andersoni in humans and a new challenge with respect to cryptosporidiosis with an increasing population of elderly people and patients with immunosuppressive diseases.
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Prevalence and Genetic Characterization of Cryptosporidium, Giardia and Enterocytozoon in Chickens From Ezhou, Hubei, China
Article
Full-text available
  • Jan 2020
Cryptosporidium spp., Giardia spp. and microsporidia are important intestinal protozoa responsible for diarrhea in humans and other mammals. China is a major chicken-raising country, and studies on these protozoa in chickens have important public health significance. Here, we investigated the prevalence and genetic characterization of these parasites in chickens from Ezhou City, Hubei Province, China. In total, 206 stool specimens were collected from chickens in four villages of Ezhou between July 2014 and February 2015. Genomic DNA of each specimen was tested by nested PCR based on the Cryptosporidium small subunit rRNA gene, the Giardia intestinalis triose phosphate isomerase gene, and the internal transcribed spacer of the Enterocytozoon bieneusi rRNA gene, respectively. The public health significance of G. intestinalis and E. bieneusi identified in our study was evaluated via phylogenetic analysis. The infection rates were determined to be 2.43% (5/206), 8.25% (17/206), and 1.94% (4/206) for Cryptosporidium, G. intestinalis, and E. bieneusi, respectively. One sample showed coinfection with G. intestinalis and E. bieneusi. Meanwhile, sequence analysis of the PCR-positive samples showed that the Cryptosporidium was C. baileyi, G. intestinalis was assemblage C, and E. bieneusi was genotype D and novel genotype EZ0008. This is the first report of zoonotic G. intestinalis assemblage C in chickens in the world, and the first report of zoonotic E. bieneusi genotype D in chickens in China. These findings indicate new transmission dynamics and molecular epizootiology.
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First Detection of Cryptosporidium spp. in Migratory Whooper Swans (Cygnus cygnus) in China
Article
Full-text available
  • Dec 2019
Cryptosporidium is an important protozoan parasite that can cause gastrointestinal diseases in humans and that also causes respiratory and gastrointestinal diseases in birds. In this study, we investigated the occurrence of Cryptosporidium species in migratory whooper swans in China. Fecal samples (n = 467) from whooper swans were collected from Sanmenxia Swan Lake National Urban Wetland Park, China. The samples were analyzed for Cryptosporidium species and genotypes with PCR along a sequence analysis of the small subunit rRNA. Cryptosporidium was detected in eight of the 467 (1.7%) samples. The analysis of the small subunit rRNA sequence data revealed two zoonotic species (Cryptosporidium parvum and Cryptosporidium andersoni) and one genotype (Cryptosporidium goose genotype II). These are the first data on the positive rate of Cryptosporidium spp. in whooper swans in China, and they suggest that whooper swans can harbor the zoonotic species C. parvum and C. andersoni in China.
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PCR-Based Detection of Cryptosporidium spp. and Enterocytozoon bieneusi in Farm-Raised and Free-Ranging Geese (Anser anser f. domestica) From Hainan Province of China: Natural Infection Rate and the Species or Genotype Distribution
Article
Full-text available
  • Dec 2019
Cryptosporidium spp. and Enterocytozoon bieneusi are two important zoonotic pathogens that can infect humans and a broad range of animal hosts. However, few studies have been conducted to study infection of the two pathogens in domestic geese until now. The aims of the present study were to determine the prevalence of natural infection, and the species or genotype distribution of Cryptosporidium and E. bieneusi in farm-raised and free-ranging geese from Hainan Province of China. In total, 266 fecal samples of geese were collected (142 farm-raised and 124 free-ranging geese). Cryptosporidium spp. and E. bieneusi were identified by nested PCR and sequencing analysis of the SSU rRNA and the ITS region of the rRNA genes. A total of 4.1% (12/226) of the geese were positive for Cryptosporidium spp., with 0.7% identified in the farm-raised geese and 7.0% in the free-ranging geese. Two bird-adapted species/genotypes were identified: C. baileyi (n = 1) and Cryptosporidium goose genotype I (n = 11). Meanwhile, E. bieneusi was found in 13.9% (37/266) of geese, with 8.9% identified in the farm-raised and 21.8% in the free-ranging geese. Eleven genotypes of E. bieneusi were identified constituted with six known genotypes: D (n = 13), I (n = 5), CHG2 (n = 1), CHG3 (n = 5), and CHG5 (n = 1), and five novel genotypes named HNE-I to V (one each). All of the genotypes identified in the geese here belonged to zoonotic Groups 1 or 2. This study is the first to demonstrate the presence of Cryptosporidium spp. and E. bieneusi in domestic geese from Hainan, China, and provides baseline data that will be useful for controlling and preventing these pathogens in goose farms. The geese infected with E. bieneusi, but not with Cryptosporidium, should be considered potential public health threats.
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Occurrence of a Cryptosporidium xiaoi–like genotype in peafowl (Pavo cristatus) in China
Article
Full-text available
  • Nov 2019
The aim of this study was to survey the Cryptosporidium species in peafowls (Pavo cristatus) in Henan Province, China. A total of 143 fecal specimens collected from a breeding farm were tested for Cryptosporidium by nested PCR targeting the small subunit rRNA (SSU rRNA), 70-kDa heat shock protein (HSP70), and actin genes of Cryptosporidium followed by sequence analysis. Only one isolate from an asymptomatic host was obtained, and the isolate differed from a new C. xiaoi–like genotype by one nucleotide and from C. xiaoi or C. bovis at the SSU rRNA locus by six nucleotides. Likewise, the actin gene shared 99% identity with the C. xiaoi–like genotype, accompanied by four nucleotide mutations. A complete sequence of the HSP70 gene was obtained, and exhibited 96% similarity with that from C. xiaoi and differed by one nucleotide from that with the C. xiaoi–like genotype. Phylogenetic analysis of the current isolate revealed genetic relatedness to the C. xiaoi–like genotype and distinction from C. xiaoi and C. bovis. Therefore, our results provided the first documentation of avian infection with a C. xiaoi–like genotype in China and further insight into the diversity of Cryptosporidium spp. in avians.
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Molecular investigation of Cryptosporidium in farmed chickens in Hubei Province, China, identifies ‘zoonotic’ subtypes of C. meleagridis
Article
Full-text available
  • Aug 2018
Background: Cryptosporidium is a key genus of parasitic protists that infect humans and other vertebrates (mammals and birds). Birds are typically infected with C. avium, C. baileyi, C. galli and/or C. meleagridis, the latter of which is recognised as being zoonotic. Stimulated by the previous finding of C. meleagridis subtypes IIIbA21G1R1, IIIbA22G1R1 and IIIbA26G1R1 in diarrhoeic children in Wuhan city and environs in Hubei Province, China, we performed a molecular epidemiological survey to explore whether these or similar subtypes might occur in farmed chickens in this province. Methods: PCR-coupled sequencing analyses of regions in the small subunit (SSU) of the nuclear ribosomal RNA and 60 kDa glycoprotein (gp60) genes were utilised to characterise Cryptosporidium in faecal samples from chickens (n = 471) from 14 farms from six distinct regions in Hubei Province. Results: Cryptosporidium baileyi (33/471; 7.0%) and C. meleagridis (15/471; 3.2%) were identified in chickens on eight farms in five of the six distinct geographical regions. No significant age-associated difference in the prevalence of C. baileyi was evident, whereas the prevalence of C. meleagridis was significantly higher in younger (≤ 4 months) than in older chickens (> 4 months). For C. meleagridis, two subtype families, IIIb and IIIe, were defined; some of the subtypes (i.e. IIIbA26G1R1b and IIIbA22G1R1c) characterised here matched those identified previously in diarrhoeic children in Wuhan. Conclusions: This is the first molecular study reporting the genetic identity and prevalence of C. baileyi and C. meleagridis in chickens in Hubei. The findings suggest that C. meleagridis subtypes IIIbA26G1R1b and IIIbA22G1R1c are cross-transmissible between chickens and humans, raising awareness about the significance of birds as potential reservoirs of zoonotic variants of Cryptosporidium. Future studies might focus on investigating the prevalence of 'zoonotic' subtypes of Cryptosporidium meleagridis in various species of wild and domesticated birds, and on comparing them with those found in humans in China and other countries.
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Molecular Identification of Cryptosporidium spp., Enterocytozoon bieneusi, and Giardia duodenalis in Captive Pet Birds in Henan Province, Central China
Article
  • Jan 2021
Cryptosporidium spp., Enterocytozoon bieneusi, and Giardia duodenalis are common enteric pathogens that are capable of infecting humans and animals. Total of 1,005 fecal samples from captive pet birds were collected from seven locations in Henan Province, China. The results demonstrated that 9.9% (99/1,005) of the captive birds were infected with one of these three pathogens. E. bieneusi was the most prevalent species among the birds (45/1,005, 4.5%) followed by G. duodenalis (33/1,005, 3.3%) and Cryptosporidium spp. (21/1,005, 2.1%). Five Cryptosporidium species were identified, namely, C. baileyi (10), C. galli (5), C. meleagridis (4), C. andersoni (1), and C. parvum (1). Two known E. bieneusi genotypes were identified: Peru 6 (44) was identified in pigeons (34) and European turtle doves (10), whereas the genotype PtEb I (1) was only identified in a pigeon. Only G. duodenalis assemblage E (33) was identified in some pet birds. To the best of our knowledge, this study is the first to undertake the molecular identification of G. duodenalis in birds in China. The identification of potentially zoonotic species/genotypes of the pathogens suggests that exposure to the excreta of these birds, either directly or via food and water, may pose a threat to human health.
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A One Health Approach to Tackle Cryptosporidiosis
Article
  • Jan 2020
  • Trends Parasitol
Cryptosporidiosis is a significant diarrhoeal disease in both people and animals across the world and is caused by several species of the protozoan parasite Cryptosporidium. Recent research has highlighted the longer-term consequences of the disease for malnourished children, involving growth stunting and cognitive deficits, and significant growth and production losses for livestock. There are no vaccines currently available to prevent the disease and few treatment options in either humans or animals, which has been a significant limiting factor in disease control to date. A One Health approach to tackle zoonotic cryptosporidiosis looking at new advances in veterinary, public, and environmental health research may offer several advantages and new options to help control the disease.
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Molecular study and genotyping of Cryptosporidium baileyi and Cryptosporidium parvum from free-range and commercial broiler chickens in Guilan province, Iran
Article
  • Jan 2020
Cryptosporidiosis acutely impacts the digestive and/or respiratory tract of the birds in many species of various orders. More importantly, it is also well known as a significant zoonotic disease, which can lead to diarrhea in humans and livestock. Regarding increasing demand for free-range products and increasing the number of free-range poultry farms, the present paper evaluated histopathological and molecular detection of Cryptosporidium baileyi and Cryptosporidium parvum in free-range and commercial broiler chickens in the north part of Iran. For this purpose, 100 fecal and tissue samples of the chickens in Guilan province were collected. After microscopic examination using Ziehl-Neelsen staining, molecular analyses of the fecal samples were processed by Nested-PCR targeting the 18S rRNA gene followed by sequencing of the amplicons and phylogenetic analyses. Eventually, the tissue samples were studied for histological lesions. Findings demonstrated the presence of Cryptosporidium baileyi and Cryptosporidium parvum in 6 % and 2 % of fecal samples, respectively. This is the first identification of C.parvum in avian hosts in Iran, and for the first time, C.baileyi and C.parvum are shown in native free-range chickens in Iran. All of the PCR positive birds with clinical symptoms showed gross lesions of respiratory infections. There was no significant difference between infection rate in free-range and commercial broiler chickens; however, the infection rate was significantly higher in chickens <25 days old. To conclude, we present here a notable Cryptosporidium infection rate in the free-range chicks in Iran, which notify the role of this host as a reservoir and should be more noted due to the economic and zoonotic importance.
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